Activity of Bromelain with Cationic Surfactants and the Correlation with the Change of 1H NMR Signals

This study observed the activities of bromelain in the presence of various cationic surfactants at different temperatures and the conformational changes in bromelain by ¹H NMR spectroscopy. We found that the bromelain activity was enhanced by tens to hundreds of micromoles per liter of the surfactant. In the presence of the surfactants, bromelain exhibited good tolerance to a range of substrate temperatures and its thermal stability was also increased. The ¹H NMR experiments indicated that when the temperature was increased from 25.0 to 45.0 °C, the protons of bromelain having chemical shifts (δ) between 3.7 and 5.2 ppm moved upfield, while those having δ values between 3.2 and 3.7 ppm moved downfield. In the bromelain/cationic surfactant mixture, the values of δ for the protons in both bromelain and the cationic surfactants decreased, accompanied by the broadening of the half-peak width of the surfactant protons. These results indicated that both increasing temperature and adding a cationic surfactant made the bromelain chain more flexible and hence, increased the bromelain activity. To the best of our knowledge, this was the first time that the relationship between the protein activity and the ¹H NMR data was expounded.     

Purification Processes and Market Potential of Bromelain in Brazil

Bromelain is the denomination given to the group of endoproteases obtained from members of Bromeliacea family. These enzymes have a wide range of proven applications and have been an object of study for worldwide researchers for decades. Over the years, several different downstream processes were studied in order to determine which technique would be worthwhile to be implemented in Brazil and provide the national market with such product. The objective of the present study is to relate the main studies in Brazil that has proven that bromelain purification can be cost-effective, in addition to the well-known benefits owned by such enzymes, and highlight the applications that create their market potential in the Brazilian market.     

食品加工业中菠萝蛋白酶的热稳定性

运用动力学的方法研究了菠萝蛋白酶的热稳定性，发现某些多羟基、多羟基化合物能明星延长菠萝蛋白酶在较高温度下的半寿期。这些物质无毒无害，能安全有效的提高食品加工业中菠萝蛋白酶的贮藏期和使用期。     

Green approach towards hydrolysing wheat gluten using waste ingredients from pineapple processing industries

In this work, wheat gluten was hydrolysed with pineapple crude extract, obtained from pineapple core which is a waste from the pineapple processing industry. The efficacy of crude pineapple extract was at par with the commercial bromelain in terms of hydrolysing wheat gluten. The hydrolysis proceeded at a rapid rate during the first 60 min, then slowed down till 120 min, followed by a further steep rise until 180 min and a very slow reaction up to the 24-h incubation period. The resulting hydrolysates had a fair solubility (>40%) over a pH range of 2–10. The molecular masses of the identified mixture of peptides ranged between 500 and 5000 Da, with 16% and 79% of the identified peptides in the range 0.5–1.0 and 1.0–2.0 kDa, respectively. The functional properties of the hydrolysates were remarkably better than the original gluten.     

Peroral Administration of Enzymes: Strategies to Improve the Galenic of Dosage Forms for Trypsin and Bromelain

In this study, we investigated the presystemic metabolism of trypsin and bromelain and the influence of these proteolytic enzymes on the mucus layer covering the gastrointestinal (GI) epithelia. In vitro studies demonstrated that 77.3% ± 4.0% (mean ± SD, n = 3) of trypsin is autodegraded within 2 hr, whereas autodegradation of bromelain was negligible. In contrast to the metabolization of bromelain by all pancreatic serine proteases, trypsin is only degraded to some extent by elastase. Both therapeutically used enzymes remained stable after incubation with an excised porcine mucosa, demonstrating that proteolysis caused by brush border membrane–bound enzymes is negligible. Trypsin and bromelain were highly mucolytic active, thereby reducing the diffusion barrier based on the mucus gel layer. Strategies to improve the galenic of dosage forms for trypsin and bromelain include the use of bioadhesive polymers such as hydroxyethylcellulose or slightly modified chitosan-EDTA, providing strongly improved stability of these enzymes toward proteolytic degradation in vitro. The given information represents a good starting point to improve the galenic of dosage forms for orally administered proteolytic enzymes.     

Release of antimicrobial peptides through bromelain hydrolysis of leatherjacket (Meuchenia sp.) insoluble proteins

Antibacterial peptides have been found to be a natural part of animal and plant defence systems. In some cases, antimicrobial peptides have been found to be released by hydrolysis of food proteins. In the present study, two antibacterial peptide fractions (fractions 9 and 12) had been isolated from bromelain hydrolysate of leatherjacket (Meuchenia sp.) insoluble muscle proteins. Assay for antimicrobial activity showed that fraction 12 had a minimum inhibition concentration (MIC) value of 4.3 mg/ml against Bacillus cereus and Staphylococcus aureus, while fraction 9 only showed some activity against Bacillus cereus without a MIC being reached at a 5.35 mg/ml peptide concentration. Further fractionation, on an analytical C-18 column, indicated that the fractions contained many other peptides that could account for the high MIC value relative to the cationic antibiotic polymyxin.     

菠萝蛋白酶的提取及其在医药中的应用

综述了菠萝蛋白酶的三种提取方法,并分析各自的特点,与高岭土吸附法和单宁沉淀法相比,超滤法能使产品质量、纯度、酶活都优于前者,是菠萝蛋白酶升级换代的新工艺。同时,综述了菠萝蛋白酶在医药中的应用,并指出对菠萝蛋白酶进行化学修饰以提高其稳定性和活力是重要的研究方向。     

Peroral administration of enzymes: strategies to improve the galenic of dosage forms for trypsin and bromelain

In this study, we investigated the presystemic metabolism of trypsin and bromelain and the influence of these proteolytic enzymes on the mucus layer covering the gastrointestinal (GI) epithelia. In vitro studies demonstrated that 77.3% ± 4.0% (mean ± SD, n = 3) of trypsin is autodegraded within 2 hr, whereas autodegradation of bromelain was negligible. In contrast to the metabolization of bromelain by all pancreatic serine proteases, trypsin is only degraded to some extent by elastase. Both therapeutically used enzymes remained stable after incubation with an excised porcine mucosa, demonstrating that proteolysis caused by brush border membrane-bound enzymes is negligible. Trypsin and bromelain were highly mucolytic active, thereby reducing the diffusion barrier based on the mucus gel layer. Strategies to improve the galenic of dosage forms for trypsin and bromelain include the use of bioadhesive polymers such as hydroxyethylcellulose or slightly modified chitosan-EDTA, providing strongly improved stability of these enzymes toward proteolytic degradation in vitro. The given information represents a good starting point to improve the galenic of dosage forms for orally administered proteolytic enzymes.     

Lipase activity and fatty acid typoselectivities of plant extracts in hydrolysis and interesterification

Lipase fatty acid typoselectivities of Euphorbia characias latex and commercially available crude preparation of bromelain were determined in the hydrolysis of homogeneous triacylglycerols (TAG) and natural TAG mixtures. Their activities were compared to a commercially available crude preparation of papain. Under optimal lipolysis conditions at pH 8.0 and 10 min of incubation time, maximal activities were observed at 45, 55, and 50°C, respectively, for E. characias latex, crude bromelain, and crude papain. Commercially available crude preparations of bromelain exhibited very poor hydrolysis activity. Latex from E. characias, which contained 340 mg of dried material per milliliter of fresh latex, exhibited a high lipase activity and a short-chain fatty acid preference in the hydrolysis of homogeneous TAG. For all substrates, it showed a better activity than crude papain. Lipase fatty acid typoselectivities of crude bromelain and crude papain also were studied in interesterification reactions of tributyrin with a series of homogeneous TAG. Experiments showed that crude bromelain [water activity (A _w )∶ 0.21] had no activity in interesterification. Regarding reactions with crude papain (A _w ∶ 0.55), yields of newly formed TAG decreased with increasing chain length of TAG, except for the reaction with trimargarin. For interesterification of tributyrin with unsaturated TAG, triolein reacted faster than polyunsaturated TAG. During these interesterification reactions, the proportion of new TAG with two butyroyl residues was higher than new TAG with only one butyroyl residue. This phenomenon was more pronounced for reactions with long-chain TAG.